Chloramphenicol ELISA 试剂盒
适用: 化学剂
Colorimetric
Competition ELISA
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- 抗原 See all Chloramphenicol products
- Chloramphenicol
- 适用
- 化学剂
- 检测方法
- Colorimetric
- 实验类型
- Competition ELISA
- 应用范围
- ELISA
- 原理
- Enzyme Immunoassay for the Quantitative Determination of Chloramphenicol in Food
- Analytical Method
- Quantitative
- 特异性
- 90-115%
- 灵敏度
- 0.03 ng/mL
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- 实验时间
- 1 h
- 板类型
- Pre-coated
- 实验流程
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- Prepare samples as described above.
2. Pipet 100 μL standards or prepared samples in duplicate into the appropriate wells of the microtiter plate. Immediately add 50 μL anti-chloramphenicol antibody into each well.
3. Cover the microtiter plate with a plastic foil and incubate for 30 minutes at room temperature.
4. Without preceding washing add 50 μL chloramphenicol-peroxidase conjugate into each well.
5. Cover the microtiter plate with a plastic foil and incubate additional 15 minutes at room temperature.
6. Wash the plate three times as follows: Discard the contents of the wells (dump or aspirate). Pipet 300 μL of diluted washing solution into each well. After the third repetition empty the wells again and remove residual liquid by striking the plate against a paper towel. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbencies.
7. Pipet 100 μL of substrate solution into each well.
8. Allow the reaction to develop in the dark (e.g. cupboard or drawer, the chromogen is light-sensitive) for 15 minutes at room temperature.
9. Stop enzyme reaction by adding 100 μL of stop solution (0.5 M H2SO4) into each well. The blue color will turn yellow upon addition.
10. After thorough mixing, measure absorbance at 450 nm (reference wavelength 620 nm), using an ELISA reader. The colour is stable for 30 minutes.
- Prepare samples as described above.
- 结果分析
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- Calculate the average optical density (OD 450 nm) for each set of reference standards or samples.
2. Construct a standard curve by plotting the mean optical density obtained for each reference standard against its concentration in ng/mL on semi-log graph paper with the optical density on the vertical (y) axis and the concentration on the horizontal (x) axis.
3. Using the mean optical density value for each sample, determine the corresponding concentration of chloramphenicol in ng/mL from the standard curve. Depending on experience and/or the avail ability of computer capability, other methods of data reduction may be employed.
4. The diluted samples must be further converted by the appropriate sample dilution factor. The factors are listed for each sample matrix in the sample preparation section. Note: Due to the extraction with ethyl acetate negative samples may show a certain blank value. In repetitive performed experiments with negative samples for each matrix the following blank values were identified. Milk (direct assay) < 0.1 ng/g Milk (ethyl acetate extraction) < 0.1 ng/g Honey < 0.2 ng/g Shrimps < 0.2 ng/g Meat < 0.2 ng/g Fish meal < 0.2 ng/g Whole egg < 0.05 ng/g These values are defined as the cut-off of the method for the respective matrices. Lower concentrations have to be considered as negative. TYPICAL STANDARD VALUES The following table contains an example for a typical standard curve. The binding is calculated as percent of the absorption of the 0 ng/mL standard. These values are only an example and should not be used instead of the standard curve which has to be measured in each new test. Chloramphenicol (ng/mL) % binding of 0 ng/mL 0 100 0.05 84 0.1 70 0.5 28 1 17 5 8 PERFORMANCE
- Calculate the average optical density (OD 450 nm) for each set of reference standards or samples.
- 限制
- 仅限研究用
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- 储存条件
- 4 °C
- 储存方法
- Store at 2-8 °C
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- 抗原 See all Chloramphenicol products
- Chloramphenicol
- Abstract
- Chloramphenicol 产品
- 物质类
- Chemical
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